CN109810925A - A kind of improved polyoxin fermentation medium and zymotechnique - Google Patents

A kind of improved polyoxin fermentation medium and zymotechnique Download PDF

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Publication number
CN109810925A
CN109810925A CN201910203044.7A CN201910203044A CN109810925A CN 109810925 A CN109810925 A CN 109810925A CN 201910203044 A CN201910203044 A CN 201910203044A CN 109810925 A CN109810925 A CN 109810925A
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fermentation medium
polyoxin
zymotechnique
formula
fermentation
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CN109810925B (en
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杨玉旺
潘忠成
李昶志
李蒲民
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Shaanxi Microbe Bio-Technology Co Ltd
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Shaanxi Microbe Bio-Technology Co Ltd
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    • YGENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y02TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
    • Y02PCLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
    • Y02P60/00Technologies relating to agriculture, livestock or agroalimentary industries
    • Y02P60/80Food processing, e.g. use of renewable energies or variable speed drives in handling, conveying or stacking
    • Y02P60/87Re-use of by-products of food processing for fodder production

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Abstract

The present invention relates to a kind of improved polyoxin fermentation medium and zymotechnique, the polyoxin fermentation medium is made of formula as below: corn flour 1-2%, soybean cake powder 1.5-3%, maltose 1.5-3%, KH2PO4For 0.05-0.2%, NaCl0.05-0.2%, CaCO3For 0.15-0.45%, defoaming agent 0.025-0.050%, (NH4)2SO40.1-2%, surplus are aqua sterilisa.The present invention optimizes zymotechnique, improves the stability of fermentation medium, also improves the biological value of zymotechnique.

Description

A kind of improved polyoxin fermentation medium and zymotechnique
Technical field
The invention belongs to technical field of microbial fermentation, and in particular to a kind of improved polyoxin fermentation medium and hair Ferment technique.
Background technique
Polyoxin (Polyoxin) has preferable uptake and translocation effect, belongs to broad-spectrum antibiotic fungicide, is The safety pesticide of a kind of high-efficiency low-toxicity, non-environmental-pollution, so being widely used in the important disease such as cereal crops, fruits and vegetables Harmful prevention and treatment.It is mainly used for preventing and treating alternaria leaf spot of apple, rice sheath blight disease, ring spot, Pear black spot, grape grey mould, frost The ten various crop disease such as mildew, black fleck disease of ginseng and Alternaria alternate.
Existing polyoxin raw medicine mostly uses golden streptomyces chromogenes No4896 wild type original seed to obtain through microbial fermentation , but fermentation unit is not high and unstable.The process flow of bacterium fermentation polyoxin are as follows: mini number.
Existing polyoxin zymotechnique there is a problem of following: 1, maintaining under basic pH environment, cause more for a long time Antimycin is unstable under alkaline environment;2, the elimination of foam is gone out in existing culture medium high pressure when the viscosity and sterilizing of culture medium It cannot scientifically be controlled when bacterium.
Chinese patent application 201410624720.5 discloses a kind of polyoxin fermentation medium, the fermentation medium Contain: the cornstarch 8-11g/L of activation processing, beancake powder 18-22g/L, yeast powder 4-6g/L, sodium chloride 4-6g/L, sulfuric acid Ammonium 0.8-1.2g/L, calcium carbonate 1.8-2.2g/L;Wherein, it is 0.08- that the cornstarch of the activation processing, which is with concentration, The aqueous slkali or concentration of 0.12mol/L is that the acid solution of 0.08-0.12mol/L carries out cornstarch to embathe acquisition.Benefit With fermentation process provided by the present invention, fermentation strain is enabled to improve the utilization rate of fermentation medium, utilization rate can be with Reach 80% or more, can averagely shorten the fermentation time of about 5h, significantly improves the content of polyoxin in fermentation clear liquid, generally , the potency of polyoxin is not less than 26000 μ g/mL in fermentation clear liquid.
But the above prior art does not specialize in the quality stable problem of everfermentation culture medium.It is mostly anti-in order to improve The quality stability and confecting efficiency of mycin fermentation medium solve the problems, such as such current culture medium unstable quality, improve The production potency of fermentation reduces ferment filtrate processing difficulty, seeks a kind of improved polyoxin fermentation medium and work of fermenting Skill has great importance.
Summary of the invention
In order to solve the above-mentioned problems in the prior art, the present invention provides a kind of improved polyoxin fermented and cultured Base and zymotechnique, optimize zymotechnique, improve the stability of fermentation medium, also improve the biology of zymotechnique Potency.
The present invention adopts the following technical scheme: a kind of improved polyoxin fermentation medium, is made of formula as below: beautiful Rice flour 1-2%, soybean cake powder 1.5-3%, maltose 1.5-3%, KH2PO4For 0.05-0.2%, NaCl0.05-0.2%, CaCO3For 0.15-0.45%, defoaming agent 0.025-0.050%, (NH4)2SO40.1-2%, surplus are aqua sterilisa.
In the preferred embodiment of the present invention, the polyoxin fermentation medium is made of formula as below: corn Powder 1.5%, soybean cake powder 2.0%, maltose 2.0%, KH2PO4 0.1%、NaCl 0.1%、CaCO30.3%, defoaming agent 0.035%, (NH4)2SO40.1-2%, surplus are aqua sterilisa.
In the preferred embodiment of the present invention, the polyoxin fermentation medium is prepared via a method which It arrives, proportionally weighs each component in formula, pH 6.5 before medium sterilization, then sterilizing obtains.
In the preferred embodiment of the present invention, the sterilizing is the 30-60min that sterilizes at 120-130 DEG C.
The present invention also protects the application based on above-mentioned fermentation medium in zymotechnique, and the zymotechnique is by type Number for 4896 golden streptomyces chromogenes be inoculated into seed culture fluid after, cultivate 20-50h at 28 DEG C, culture transferring is to sterilized The polyoxin fermentation medium relays supervention ferment, presses 0.045-0.06MPa in 28-30 DEG C, 100-120rpm, tank, leads to When air quantity 0.5-1.0L/min is cultivated to 4-12h, (NH is added in feed supplement4)2SO4And ammonium hydroxide, maintain 6.3-6.5's adjusting pH Nitrogen source is supplemented simultaneously, puts tank when cultivating 110-130h.
In the preferred embodiment of the present invention, the addition (NH4)2SO4Mass concentration is 1%-5%.
Compared with prior art, the beneficial effects of the present invention are:
(1) it is obtained using golden streptomyces chromogenes fermentation medium and its improved zymotechnique, fermentation period of the invention It is effective to shorten, greatly improve fermentation efficiency.
(2) ventilation quantity also further decreases in zymotechnique simultaneously, greatly reduces the demand to air, energy source and power disappears Consumption reduces, and has saved cost.
(3) the more traditional zymotechnique of mycelia is longer, and faster, mycelial concentration improves for growth, after 10 batches of production potency comparisons, 662U/mL(18.5% is still improved using the biological value of fermentation medium of the present invention zymotechnique of the present invention).
Detailed description of the invention
It is described further with reference to the accompanying drawing:
Fig. 1 is mycelia figure of the comparative example 2 in 30h;
Fig. 2 is mycelia figure of the embodiment 2 in 30h;
Fig. 3 is mycelia figure of the comparative example 2 in 68h;
Fig. 4 is mycelia figure of the embodiment 2 in 68h;
Fig. 5 is mycelia figure of the comparative example 2 when putting tank;
Fig. 6 is mycelia figure of the embodiment 2 when putting tank;
Note: mycelia observation is to observe to obtain with 100 times of oil mirrors after the dyeing of pinkish red dye liquor.
Specific embodiment
In order to which the purpose of the present invention, technical solution and technical effect are more clearly understood, attached drawing and tool are now compareed The present invention is described in more detail for body embodiment.
Comparative example 1
Former fermentative medium formula: former fermentative medium formula: dehydrated potato powder 1.5%, soybean cake powder 2.0%, maltose 2.0%, KH2PO4 0.1%、NaCl 0.1%、CaCO3 0.3%、(NH4)2SO40.5%, defoaming agent 0.035%, surplus are aqua sterilisa.
Comparative example 2
The fermentation medium of comparative example 1 is as follows using the primary ferment technique of former fermentation medium: (slant pore → seed flask → First class seed pot (tank holds 1m3) → fermentor (tank holds 35m3) → acidification (oxalic acid) → filtering → concentration → refined filtration → spraying dry It is dry.
Embodiment 1
Fermentation medium of the invention: corn flour 1.5%, soybean cake powder 2.0%, maltose 2.0%, KH are weighed according to formula2PO4 0.1%、NaCl 0.1%、CaCO3 0.3%、(NH4)2SO40.5%, defoaming agent 0.035%, adjusting pH is 6.5 before the big gun agent that disappears is added, 121 DEG C of sterilizing 30min, obtain fermentation medium of the invention.
Embodiment 2
The fermentation medium of embodiment 1 is as follows using zymotechnique of the invention: by the golden streptomyces chromogenes of model 4896 After slant pore is inoculated into 28 DEG C of seed flask, 220rpm shaken cultivation 30h, and bottle is at shake-flask seed.Shake-flask seed is inoculated into 28 DEG C of first class seed pot culture for 24 hours, in the fermentation medium that culture transferring is prepared to sterilized embodiment 1, in 28 DEG C, 110rpm, tank pressure 0.05MPa, ventilation quantity 0.5-1.0L/min continue to ferment.(NH is added in feed supplement when culture is to 10h4)2SO4And ammonia Water supplements nitrogen source while adjusting pH and maintaining 6.3-6.5, puts tank when cultivating 113h.
Embodiment 3
The experimental result of comparative example 1 and 2 and Examples 1 and 2 is compared as follows:
Fig. 1 is using the primary ferment technique of former fermentation medium, i.e. comparative example 2, the figure of the mycelia in 30h, it is seen then that its formation Small net, mycelia are long and few.Fig. 2 is the zymotechnique using fermentation medium of the invention, i.e. embodiment 2, the bacterium in 30h The figure of silk, it is seen then that net is to big net, mycelia length and complex cross-linked in its formation, and mycelia is few, and dyeing is deep.
Fig. 3 is using the primary ferment technique of former fermentation medium, i.e. comparative example 2, the figure of the mycelia in 68h, it is seen then that its Form big net, mycelia length and complex cross-linked.Fig. 4 is the zymotechnique using fermentation medium of the invention, i.e. embodiment 2, The figure of mycelia when 68h, it is seen then that it is also to form big net, mycelia length and complex cross-linked.
Fig. 5 is using the primary ferment technique of former fermentation medium, i.e. comparative example 2 can in the figure for putting the mycelia of tank constantly See, self-dissolving is serious, and dyeing is uneven or even fuzzy and shallow, and mycelia is thin.Fig. 6 is the fermentation work using fermentation medium of the invention Skill, i.e. embodiment 2, the figure of the mycelia when putting tank, it is seen then that its even dyeing, end slightly self-dissolving, dyeing is deep, and mycelia is thick.
In addition, 10 batches of mean titres of comparative example 2 are that 3569U/ml(tests shaking flask potency), and 10 criticize flats of embodiment 2 Equal potency is 4231U/ml.
As it can be seen that using golden streptomyces chromogenes fermentation medium of the invention and its improved zymotechnique, fermentation period Current 113h is foreshortened to, fermentation period shortens 30% or more, and it is calculated within 300 days by annual production, can increase by about 21 tanks every year, Improve the efficiency of fermentation.Ventilation quantity is reduced to current 0.5-1.0L/min, greatly reduces the demand to air, and the energy is dynamic Power consumption reduces.Mycelia is longer than before, and faster, mycelial concentration improves for growth, after 10 batches of production potency comparisons, using the present invention The biological value of fermentation medium zymotechnique of the present invention improves 662U/mL(18.5%).
Although above having used general explanation, specific embodiment and test, the present invention is made to retouch in detail It states, but on the basis of the present invention, it can be made some modifications or improvements, this is apparent to those skilled in the art 's.Therefore, these modifications or improvements without departing from theon the basis of the spirit of the present invention, belong to claimed Range.

Claims (7)

1. a kind of improved polyoxin fermentation medium, which is characterized in that be made of formula as below: corn flour 1-2%, soya bean Cake powder 1.5-3%, maltose 1.5-3%, KH2PO4For 0.05-0.2%, NaCl0.05-0.2%, CaCO3For 0.15-0.45%, defoaming agent For 0.025-0.050%, (NH4)2SO40.1-2%, surplus are aqua sterilisa.
2. polyoxin fermentation medium according to claim 1, which is characterized in that be made of formula as below: corn flour 1.5%, soybean cake powder 2.0%, maltose 2.0%, KH2PO4 0.1%、NaCl 0.1%、CaCO30.3%, defoaming agent 0.035%, (NH4)2SO40.1-2%, surplus are aqua sterilisa.
3. polyoxin fermentation medium according to claim 1, which is characterized in that the polyoxin fermented and cultured Base is prepared via a method which to obtain, and proportionally weighs each component in formula, pH6.5 before disappearing, then sterilizing obtains.
4. polyoxin fermentation medium according to claim 3, which is characterized in that the sterilizing is in 120-130 Sterilize 30-60min at DEG C.
5. application of the polyoxin fermentation medium in zymotechnique described in any one of -4 according to claim 1, special Sign is that the technique is after the golden streptomyces chromogenes of model 4896 are inoculated into seed culture fluid, to train at 28 DEG C 20-50h is supported, culture transferring to the sterilized polyoxin fermentation medium relays supervention ferment, in 28-30 DEG C, 100- When 120rpm, tank pressure 0.045-0.06MPa, ventilation quantity 0.5-1.0L/min are cultivated to 4-12h, (NH is added in feed supplement4)2SO4And ammonia Water, then adjust and supplement nitrogen source while pH maintains 6.3-6.5, tank is put when cultivating 110-130h.
6. application according to claim 5, which is characterized in that be in 28 DEG C, 110rpm, tank pressure 0.05MPa, ventilation quantity When 0.5-1.0 L/min is cultivated to 4-12h, (NH is added in feed supplement4)2SO4And ammonium hydroxide.
7. application according to claim 5, which is characterized in that the addition (NH4)2SO4Mass concentration is 1%-5%.
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Publication number Priority date Publication date Assignee Title
CN110656141A (en) * 2019-10-14 2020-01-07 陕西麦可罗生物科技有限公司 Fermentation process of polyoxin for preventing and treating panax notoginseng black spot
CN110656141B (en) * 2019-10-14 2023-01-13 陕西麦可罗生物科技有限公司 Fermentation process of polyoxin for preventing and treating panax notoginseng black spot

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